Keratocyte Loss after Different Methods of De-epithelialization

Purpose: To evaluate the response of corneal stromal cells to different types of superficial injury. Methods: Twenty-two rabbits were randomized into five groups of four (with 2 untreated controls), and their corneas de-epithelialized (1) with a blunt instrument alone; (2) with an instrument and application of 100% ethanol, 0.5% proparacaine, or 4% cocaine; or (3) with the excimer laser. Twenty-four hours after surgery, the eyes were enucleated, and histologic changes were quantitated. Results: All the methods of de-epithelialization used resulted in a decrease in the number of keratocytes relative to the control numbers (P = 0.0001). There is a significantly greater decrease in keratocyte counts with 0.5% proparacaine and 100% ethanol when compared with eyes injured by mechanical means, with 4% cocaine, or with the excimer laser (P = 0.009). All treatment groups showed more polymorphonuclear leukocytes than did controls (P < 0.0001). Mechanical de-epithelialization alone or in conjunction with proparacaine produced the least inflammatory response, but de-epithelialization with the laser was associated with a greater inflammatory response (P < 0.0001). Conclusions: All methods of de-epithelialization produced a significant decrease in rabbit cornea stromal keratocytes 24 hours after injury, associated with acute inflammation. Thus, it may be appropriate to avoid using chemicals, or if chemicals are used, to at least avoid using 100% ethanol. The applicability of these findings to humans has not yet been established.